OR15-5 Human Sex Determination at the Edge of Ambiguity: Impaired SRY Phosphorylation Attenuates Expression of the Male Program

A paradox is posed by metazoan gene-regulatory networks (GRNs) that are robust yet evolvable. Insight may be obtained through studies of bistable genetic circuits mediating developmental decisions. A model in organogenesis is provided by the sex-specific differentiation of the embryonic gonadal ridge to form a testis or ovary. Here, we investigated a Swyer mutation in human testis-determining factor SRY that impairs its phosphorylation in association with variable developmental outcomes: fertile male, intersex, or infertile female (46, XY pure gonadal dysgenesis). The mutation (R30I) abrogates serine phosphorylation within a putative target site for protein kinase A (PKA) N-terminal to the HMG box. Diverse processes can be regulated by protein phosphorylation, including DNA recognition by transcription factors (TFs). Phosphorylation of this site in human SRY (LRRSSSFLCT; italics) in vitro was previously shown to enhance specific DNA affinity. Biological consequences of the mutation were evaluated in SRY-responsive mammalian cell lines following transient transfection. The mutation attenuated in concert occupancy of a target enhancer (TESCO) and SOX9 transcriptional activation. These perturbations were mitigated by acidic substitution (LRIDDDFL) whereas Ala substitutions (RRAAAFL or RIAAAFL) attenuated activity to an extent similar to R30I alone. No differences were observed in nuclear localization. Mutagenesis suggested that the central Ser is most efficiently phosphorylated in accord with PKA targeting rules. Replacement of the native site by an optimized “Kemptide” PKA site (LRRASLGCT) enhanced both SRY phosphorylation and SOX9 transcriptional activation whereas a “swapped” protein-kinase C determinant (LRRSSFRRCT) blocked phosphorylation. Among SRY variants, extent of cellular phosphorylation mirrored relative in vitro efficiencies of synthetic SRY-derived peptides as PKA-specific substrates. Although several kinases are predicted in silico to target this tri-serine motif, cell-based studies implicate PKA as the relevant kinase in vivo. Our results provide evidence that primate Sry requires its phosphorylation for full gene-regulatory activity. A PKA site N-terminal to the SRY HMG box, unique to primates, exemplifies network “tinkering” through recruitment of a new regulatory linkage. Molecular characterization of the R30I inherited Swyer mutation in SRY thus demonstrates that impaired TF phosphorylation can attenuate a human developmental switch at the edge of ambiguity

Solar Cell Optical (LASER) Communication

Presented at the Miltary Wireless Communications Conferenc

Enhancing climate change communication: strategies for profiling and targeting Australian interpretive communities

Abstracthis research aimed to provide practical information about how to design communications on climate change adaptation and target these to the Australian population.   This was achieved by: (1) identifying and increasing awareness of different climate change audiences in Australia, and (2) evaluating how each audience responds to different types of climate change messages. Phase 1 of the study used audience segmentation analysis to identify the main climate change interpretive communities within Australia; that is, groups of Australians who share similar views and understandings about climate change.   A nationwide sample consisting of 3,096 Australian residents (aged 15 to 108 years, 47% male and 53% female) completed an online survey assessing a broad range of psychological and behavioural factors related to climate change.   Latent profile analysis applied to the psychological variables suggested that this Australian sample consists of five distinct interpretive communities: Alarmed (26%), Concerned (39%), Uncertain (14%), Doubtful (12%), and Dismissive (9%). Validation analyses revealed that these groups differed in terms of how they responded to perceived climate change threats, and also in their support for particular climate change mitigation and adaptation policies.   Phase 2 of the project examined how Australian interpretive communities respond to climate change adaptation messages and identified the specific message attributes that drive these responses. 1,031 Australian residents (aged 18 to 66 years, 49.8% male, 50.2% female) completed an online survey assessing a similar set of psychological and behavioural responses to climate change to those assessed in Phase 1.   Respondents subsequently viewed six climate change adaptation messages that were randomly allocated from a pool of 60 messages sourced from the internet.   Messages were pre-coded on 10 communication cues (e.g., language complexity, normative influence), and respondents rated them on four judgement dimensions: perceived threat, perceived efficacy, fear control (message rejection), and danger control (message acceptance).   Latent profile analysis applied to the psychological variables identified three climate change interpretive communities in this sample: Alarmed (34.4%), Uncommitted (45.2%), and Dismissive (20.3%).   Judgement analysis methodology (Cooksey, 1996) found that the three interpretive communities based their threat and efficacy evaluations on unique combinations of communication cues, and that high perceived threat and high perceived efficacy were related to message acceptance for all communities.   Effective messages for Dismissive respondents used simple language and did not emphasise descriptive social norms.   Uncommitted audience members responded positively to messages that focused on preventing losses and had a strong emotional component.   Alarmed respondents preferred messages that focused on local issues and had a collectivist frame. Providing specific adaptation advice in messages was found to be effective for all communities. The results largely support the Extended Parallel Processing Model of risk communication (Witte, 1992), and suggest that message attributes should be adjusted to effectively communicate with different climate change interpretive communities within Australia.Please cite this report as:Hine, D, Phillips, W, Reser, J, Cooksey, R, Marks, A, Nunn, P, Watt, S, Ellul, M 2013 Enhancing climate change communication: Strategies for profiling and targeting Australian interpretive communities, National Climate Change Adaptation Research Facility, Gold Coast, pp. 95.his research aimed to provide practical information about how to design communications on climate change adaptation and target these to the Australian population.   This was achieved by: (1) identifying and increasing awareness of different climate change audiences in Australia, and (2) evaluating how each audience responds to different types of climate change messages. Phase 1 of the study used audience segmentation analysis to identify the main climate change interpretive communities within Australia; that is, groups of Australians who share similar views and understandings about climate change.   A nationwide sample consisting of 3,096 Australian residents (aged 15 to 108 years, 47% male and 53% female) completed an online survey assessing a broad range of psychological and behavioural factors related to climate change.   Latent profile analysis applied to the psychological variables suggested that this Australian sample consists of five distinct interpretive communities: Alarmed (26%), Concerned (39%), Uncertain (14%), Doubtful (12%), and Dismissive (9%). Validation analyses revealed that these groups differed in terms of how they responded to perceived climate change threats, and also in their support for particular climate change mitigation and adaptation policies.   Phase 2 of the project examined how Australian interpretive communities respond to climate change adaptation messages and identified the specific message attributes that drive these responses. 1,031 Australian residents (aged 18 to 66 years, 49.8% male, 50.2% female) completed an online survey assessing a similar set of psychological and behavioural responses to climate change to those assessed in Phase 1.   Respondents subsequently viewed six climate change adaptation messages that were randomly allocated from a pool of 60 messages sourced from the internet.   Messages were pre-coded on 10 communication cues (e.g., language complexity, normative influence), and respondents rated them on four judgement dimensions: perceived threat, perceived efficacy, fear control (message rejection), and danger control (message acceptance).   Latent profile analysis applied to the psychological variables identified three climate change interpretive communities in this sample: Alarmed (34.4%), Uncommitted (45.2%), and Dismissive (20.3%).   Judgement analysis methodology (Cooksey, 1996) found that the three interpretive communities based their threat and efficacy evaluations on unique combinations of communication cues, and that high perceived threat and high perceived efficacy were related to message acceptance for all communities.   Effective messages for Dismissive respondents used simple language and did not emphasise descriptive social norms.   Uncommitted audience members responded positively to messages that focused on preventing losses and had a strong emotional component.   Alarmed respondents preferred messages that focused on local issues and had a collectivist frame. Providing specific adaptation advice in messages was found to be effective for all communities. The results largely support the Extended Parallel Processing Model of risk communication (Witte, 1992), and suggest that message attributes should be adjusted to effectively communicate with different climate change interpretive communities within Australia

Calibrated Tree Priors for Relaxed Phylogenetics and Divergence Time Estimation

The use of fossil evidence to calibrate divergence time estimation has a long history. More recently Bayesian MCMC has become the dominant method of divergence time estimation and fossil evidence has been re-interpreted as the specification of prior distributions on the divergence times of calibration nodes. These so-called "soft calibrations" have become widely used but the statistical properties of calibrated tree priors in a Bayesian setting has not been carefully investigated. Here we clarify that calibration densities, such as those defined in BEAST 1.5, do not represent the marginal prior distribution of the calibration node. We illustrate this with a number of analytical results on small trees. We also describe an alternative construction for a calibrated Yule prior on trees that allows direct specification of the marginal prior distribution of the calibrated divergence time, with or without the restriction of monophyly. This method requires the computation of the Yule prior conditional on the height of the divergence being calibrated. Unfortunately, a practical solution for multiple calibrations remains elusive. Our results suggest that direct estimation of the prior induced by specifying multiple calibration densities should be a prerequisite of any divergence time dating analysis

Comparative Analysis of Root Microbiomes of Rice Cultivars with High and Low Methane Emissions Reveals Differences in Abundance of Methanogenic Archaea and Putative Upstream Fermenters.

Rice cultivation worldwide accounts for ∼7 to 17% of global methane emissions. Methane cycling in rice paddies is a microbial process not only involving methane producers (methanogens) and methane metabolizers (methanotrophs) but also other microbial taxa that affect upstream processes related to methane metabolism. Rice cultivars vary in their rates of methane emissions, but the influence of rice genotypes on methane cycling microbiota has been poorly characterized. Here, we profiled the rhizosphere, rhizoplane, and endosphere microbiomes of a high-methane-emitting cultivar (Sabine) and a low-methane-emitting cultivar (CLXL745) throughout the growing season to identify variations in the archaeal and bacterial communities relating to methane emissions. The rhizosphere of the high-emitting cultivar was enriched in methanogens compared to that in the low emitter, whereas the relative abundances of methanotrophs between the cultivars were not significantly different. Further analysis of cultivar-sensitive taxa identified families enriched in the high emitter that are associated with methanogenesis-related processes. The high emitter had greater relative abundances of sulfate-reducing and iron-reducing taxa which peak earlier in the season than methanogens and are necessary to lower soil oxidation reduction potential before methanogenesis can occur. The high emitter also had a greater abundance of fermentative taxa which produce methanogenesis precursors (acetate, CO2, and H2). Furthermore, the high emitter was enriched in taxa related to acetogenesis which compete with methanogens for CO2 and H2 These taxa were enriched in a spatio-specific manner and reveal a complex network of microbial interactions on which plant genotype-dependent factors can act to affect methanogenesis and methane emissions.IMPORTANCE Rice cultivation is a major source of anthropogenic emissions of methane, a greenhouse gas with a potentially severe impact on climate change. Emission variation between rice cultivars suggests the feasibility of breeding low-emission rice, but there is a limited understanding of how genotypes affect the microbiota involved in methane cycling. Here, we show that the root microbiome of the high-emitting cultivar is enriched both in methanogens and in taxa associated with fermentation, iron, and sulfate reduction and acetogenesis, processes that support methanogenesis. Understanding how cultivars affect microbes with methanogenesis-related functions is vital for understanding the genetic basis for methane emission in rice and can aid in the development of breeding programs that reduce the environmental impact of rice cultivation

Soil domestication by rice cultivation results in plant-soil feedback through shifts in soil microbiota.

BackgroundSoils are a key component of agricultural productivity, and soil microbiota determine the availability of many essential plant nutrients. Agricultural domestication of soils, that is, the conversion of previously uncultivated soils to a cultivated state, is frequently accompanied by intensive monoculture, especially in the developing world. However, there is limited understanding of how continuous cultivation alters the structure of prokaryotic soil microbiota after soil domestication, including to what extent crop plants impact soil microbiota composition, and how changes in microbiota composition arising from cultivation affect crop performance.ResultsWe show here that continuous monoculture (> 8 growing seasons) of the major food crop rice under flooded conditions is associated with a pronounced shift in soil bacterial and archaeal microbiota structure towards a more consistent composition, thereby domesticating microbiota of previously uncultivated sites. Aside from the potential effects of agricultural cultivation practices, we provide evidence that rice plants themselves are important drivers of the domestication process, acting through selective enrichment of specific taxa, including methanogenic archaea, in their rhizosphere that differ from those of native plants growing in the same environment. Furthermore, we find that microbiota from soils domesticated by rice cultivation contribute to plant-soil feedback, by imparting a negative effect on rice seedling vigor.ConclusionsSoil domestication through continuous monoculture cultivation of rice results in compositional changes in the soil microbiota, which are in part driven by the rice plants. The consequences include a negative impact on plant performance and increases in greenhouse gas emitting microbes